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Purification of Hsp104, a Protein Disaggregase

Purification of Hsp104, a Protein Disaggregase

Article DOI: 10.3791/3190
September 30th, 2011


Summary September 30th, 2011

Here, we describe a protocol for the purification of highly active Hsp104, a hexameric AAA+ protein from yeast, which couples ATP hydrolysis to protein disaggregation. This scheme exploits a His6-tagged construct for affinity purification from E. coli followed by anion-exchange chromatography, His6-tag removal with TEV protease, and size-exclusion chromatography.

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